The ING5 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ING5 tumor suppressor gene in the HT29 human colorectal adenocarcinoma cell line. Through CRISPR/Cas9-mediated gene disruption, this polyclonal population provides a heterogeneous loss-of-function model that avoids the artifacts of single-cell cloning. This approach preserves natural genetic variability, enabling assessment of dominant signaling effects in a colorectal cancer-relevant background.
The parental HT29 cell line is an epithelial line originally isolated from a primary colorectal adenocarcinoma. It is widely employed for studying intestinal epithelial biology, drug metabolism, and cancer, and can undergo enterocytic differentiation under appropriate conditions. HT29 cells harbor endogenous TP53 mutations, reflecting the genetic landscape of many colorectal tumors and providing a pathologically relevant system for investigating tumor suppressor networks.
ING5 functions as a type II tumor suppressor and is a core component of the NuA4/TIP60 histone acetyltransferase (HAT) complex. It directly interacts with p53 and the transcriptional coactivators p300/CBP, thereby enhancing p53-mediated transcriptional activation of target genes such as CDKN1A (p21) and BAX. This activity promotes cell cycle arrest and apoptosis in response to upstream signals, including DNA damage and cellular stress. ING5 also associates with 14-3-3 proteins, which modulate its subcellular localization. Through its HAT activity, ING5 links chromatin remodeling to tumor suppression.
In the HT29 colorectal adenocarcinoma context, disruption of ING5 is predicted to compromise p53-mediated apoptotic and cell cycle arrest programs, even in the presence of mutant p53 (R273H), by affecting residual p53 functions or p53-independent roles. The polyclonal knockout population mimics tumor heterogeneity, enabling studies into how ING5 loss cooperates with pre-existing oncogenic mutations. This makes the model invaluable for dissecting the contribution of ING5 inactivation to colorectal tumorigenesis.
Key research applications include mechanistic dissection of tumor suppression, analysis of p53 pathway dynamics, and investigation of chromatin modification. Representative assays encompass Western blotting for ING5, p21, and BAX; RT-qPCR for downstream transcripts; annexin V-based apoptosis assessment; cell cycle profiling; and proliferation assays. Co-immunoprecipitation can examine ING5 interactions with p53 or HAT complex members, and ChIP-qPCR can evaluate histone acetylation at specific genomic loci. The cells are also well-suited for drug sensitivity screens and functional genomics studies. For further technical information, please contact Ascent Research.
